MAP7 regulates axon morphogenesis by recruiting kinesin-1 to microtubules and modulating organelle transport.

Neuronal cell morphogenesis depends on proper regulation of microtubule-based transport, but the underlying mechanisms are not well understood. Here, we report our study of MAP7, a unique microtubule-associated protein that interacts with both microtubules and the motor protein kinesin-1. Structure-function analysis in rat embryonic sensory neurons shows that the kinesin-1 interacting domain in MAP7 is required for axon and branch growth but not for branch formation. Also, two unique microtubule binding sites are found in MAP7 that have distinct dissociation kinetics and are both required for branch formation. Furthermore, MAP7 recruits kinesin-1 dynamically to microtubules, leading to alterations in organelle transport behaviors, particularly pause/speed switching. As MAP7 is localized to branch sites, our results suggest a novel mechanism mediated by the dual interactions of MAP7 with microtubules and kinesin-1 in the precise control of microtubule-based transport during axon morphogenesis

Spatiotemporal Control of Human Cardiac Tissue Through Optogenetics

Cardiac arrhythmias are caused by disordered propagation of electrical activity. Progress in understanding and controlling arrhythmias requires novel methods to characterize and control the spatiotemporal propagation of electrical activity. We used patterned illumination of cardiomyocytes derived from optogenetic human induced pluripotent stem cells to create dynamic conduction blocks, and to test spatially extended control schemes. Using this model, we demonstrated the ability to initiate, circumscribe, relocate, and terminate pathologic spiral waves that drive many arrhythmias. When cells were derived from patients with long QT syndrome, longer action potential durations made spiral waves more resistant to termination. This work lays the foundation for personalized models of cardiac injury and disease, and the development of tailored approaches to the management of arrhythmias

Fluorescence Near-Field Microscopy of DNA at Sub-10 nm Resolution

We demonstrate apertureless near-field microscopy of single molecules at sub-10 nm resolution. With a novel phase filter, near-field images of single organic fluorophores were obtained with ~sixfold improvement in the signal-to-noise ratio. The improvement allowed pairs of molecules separated by ~15 nm to be reliably and repeatedly resolved, thus demonstrating the first true Rayleigh resolution test for near-field images of single molecules. The potential of this technique for biological applications was demonstrated with an experiment that measured the helical rise of A-form DNA

Creating Online Lessons: A Faculty Development Seminar Series

The World Wide Web is being used increasingly to deliver instruction in medical education. Consequently, there is a need to train faculty in developing and implementing online instruction. We developed and implemented a seminar series to teach faculty to create educationally sound, well designed online instruction. Instruction was delivered to 15 participants via a six session seminar on developing web based lessons, supplemented with web-based instruction. First, the participants learned the basics of instructional design via a web based module. They then completed content outlines for their online lessons prior to the first seminar. Lesson development, web site development and the use of a web based instructional shell to implement the online lessons were each taught in two two hour sessions. Eight participants developed online lessons and four actually implemented them. Feedback was mostly positive, with suggestions for improvement. All eight participants who completed the series said they would recommend it to their colleagues. Because a longitudinal workshop type of seminar series requires a large amount of participant time outside of class, a six month seminar series may be too long. It is important at the beginning of the series to help participants select topics suitable for online instruction and to help them narrow their topics. We may change the attendance guidelines so faculty would attend only the session on instructional design and have their staff attend the technical sessions on web site design, HTML editing and online course delivery systems. This would better match the actual practice of faculty designing the instruction and staff developing it

The risk of misclassifying subjects within principal component based asset index.

The asset index is often used as a measure of socioeconomic status in empirical research as an explanatory variable or to control confounding. Principal component analysis (PCA) is frequently used to create the asset index. We conducted a simulation study to explore how accurately the principal component based asset index reflects the study subjects' actual poverty level, when the actual poverty level is generated by a simple factor analytic model. In the simulation study using the PC-based asset index, only 1% to 4% of subjects preserved their real position in a quintile scale of assets; between 44% to 82% of subjects were misclassified into the wrong asset quintile. If the PC-based asset index explained less than 30% of the total variance in the component variables, then we consistently observed more than 50% misclassification across quintiles of the index. The frequency of misclassification suggests that the PC-based asset index may not provide a valid measure of poverty level and should be used cautiously as a measure of socioeconomic status

Tip-Enhanced Fluorescence Microscopy at 10 Nanometer Resolution

We demonstrate unambiguously that the field enhancement near the apex of a laser-illuminated silicon tip decays according to a power law that is moderated by a single parameter characterizing the tip sharpness. Oscillating the probe in intermittent contact with a semiconductor nanocrystal strongly modulates the fluorescence excitation rate, providing robust optical contrast and enabling excellent background rejection. Laterally encoded demodulation yields images with <10 nm spatial resolution, consistent with independent measurements of tip sharpness